Ex Vivo Gene Expression in Whole Blood as a Model of Assessment of Individual Variation to Dietary Supplements

a whole blood and gene expression technology, applied in the direction of heterocyclic compound active ingredients, plant/algae/fungi/lichens ingredients, instruments, etc., can solve the problems of inability to employ clinical studies, double-blind clinical trials are no longer able to identify those dietary components, genotyping or single nucleotide polymorphism analysis is only useful, etc., to identify the potential anti-cancer or anti-autoimmune disorder effectiveness of dietary components

Inactive Publication Date: 2008-08-28
HITACHI CHEM CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

[0020]A further embodiment of the present invention provides a method of measuring the potential anti-cancer or anti-autoimmune disorder effectiveness in a mammal of a dietary component selected from the group consisting of vitamin A, vitamin C, vitamin D, vitamin E, epigallocatechin gallate, g-linoleic acids, genistein, curcumin, quercetin, aged garlic, agaricus, propolis, meshimakobu, noni extract, alkoxyglycerol, and fucoidan, comprising: exposing whole blood of the mammal to the dietary component for 4 hours or less; stimulating the exposed whole blood and unexposed whole blood of the mammal with phytohemagglutinin; after said stimulus, measuring the amount of mRNA encoding a protein selected from the group consisting of interleukin-2, interleukin-4, tumor necrosis factor alpha, and Fas ligand in blood cells of the exposed whole blood and the unexposed whole blood; comparing results of the measurement obtained in blood cells of the exposed whole blood and the unexposed whole blood; and identifying potential anti-cancer or anti-autoimmune disorder effectiveness of the dietary component based on the results of the comparison, wherein a change in the amount of the mRNA correlates with the effectiveness of the dietary component.

Problems solved by technology

However, it is difficult to employ clinical study results to design an individualized combination of dietary components for a particular individual.
If both responders and non-responders exist in a study population, and the non-responder population is substantially larger than the responder population, double-blind clinical trials are no longer capable of identifying those dietary components that can be expected to have efficacy.
Furthermore, genotyping or single nucleotide polymorphism analysis is only useful in dietary optimization once target genes and hot spots have been characterized.
However, the applicable technology is still limited.
However, there has not as yet been an effective method for accomplishing this, particularly in an ex vivo context.

Method used

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  • Ex Vivo Gene Expression in Whole Blood as a Model of Assessment of Individual Variation to Dietary Supplements

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embodiment 1

[0030]In order to assess leukocyte function in as close as possible to physiological conditions, an embodiment of the method of the invention employs whole blood, without isolating specific leukocyte populations. As anticoagulant citrate dextrose solution and ethylenediaminetetraacetic acid chelate calcium, a critical component for many biological activities (see Eggesbo et al., “LPS induced release of IL-1 beta, IL-6, IL-8 and TNF-alpha in EDTA or heparin anticoagulated whole blood from persons with high or low levels of serum HDL,” Cytokine 1996; 8: 152-60), heparin was used as an anticoagulant in this embodiment. Since mRNA transcription is an upstream event of protein synthesis and subsequent biological activities, mRNA levels are employed in an embodiment of the present invention as an indication of biological activity associated with the protein encoded by the mRNA. An embodiment of the present invention employs a method of quantitating mRNA that allows the identification of c...

embodiment 2

[0037]In this embodiment of the method of the present invention, the expression of CD32A mRNA was assessed in the leukocytes of an individual. This mRNA encodes the IgG Fc receptor, and is related to antibody-dependent cell-mediated cytotoxicity (ADCC). Dietary components that increase CD32A mRNA levels would be expected to boost an individual's ADCC activity and thus provide direct anticancer activity. Alternatively, such dietary components could enhance the efficacy of recently developed expensive monoclonal antibody-based treatments (such as trastuzumab (Herceptin) or rituximab (Rituxan)) by simultaneously increasing IgG Fc receptor mRNA levels in circulating leukocytes. The method described above was employed in measuring the CD32A mRNA levels, with the exception that no stimulating agent such as phytohemagglutinin was employed. The primer sequences are shown in Table 2 above.

[0038]The dietary supplements employed were: vitamin A (“VA”; 100 nmol / L, final concentration;), vitamin...

embodiment 3

[0041]In this embodiment, heparinized whole blood of four individuals was pre-incubated with various dietary supplements at 37° C. for 1-2 hours (at the same blood concentrations as in Embodiment 2), then stimulated with 1 Gy radiation. The blood was then incubated at 37° C. for 2 hours. The level of p21 mRNA was then assessed using the method described in Embodiment 1. The primer sequences are given in Table 2. p21 mRNA was selected as an apoptosis marker that would indicate the effect of each dietary component on the DNA damage response. As an alternative to p21, PUMA (p53 upregulated modulator of apoptosis) mRNA could be quantitated.

[0042]The results are shown in FIG. 3. In the Figure, open circles indicate values obtained without radiation stimulus, while closed circles indicate that radiation was used. Each symbol is the mean±S.D. from triplicate aliquots of 50 mL heparinized whole blood.

[0043]As shown in the figure, the individual responses to the dietary components varied wid...

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Abstract

A method is disclosed for individually tailoring the administration of dietary components such as supplements. In the method, whole blood of a mammal is exposed to a dietary component. The level of a marker mRNA linked to a disease state is measured in leukocytes after exposure to the dietary component, and in some cases after further stimulation of the exposed blood cells. By comparing the mRNA level after exposure with the value found in unexposed blood cells, it is possible to determine what the effect of the dietary component will be in the mammal. By screening blood of the mammal against a number of possible dietary components, it is possible to develop an optimized set of dietary components tailored to the specific mammal to treat or prevent a disease state.

Description

BACKGROUND OF THE INVENTION[0001]1. Field of the Invention[0002]The present invention relates to a method for tailoring the administration of dietary components such as supplements. In the method, whole blood of a mammal is exposed to a dietary component. The level of a marker mRNA linked to a disease state is measured in leukocytes after exposure to the dietary component, and in some cases after further stimulation of the exposed blood cells. By comparing the mRNA level after exposure with the value found in unexposed blood cells, it is possible to determine what the effect of the dietary component will be in the mammal. By screening blood of the mammal against a number of possible dietary components, it is possible to develop an optimized set of dietary components tailored to the specific mammal to treat or prevent a disease state.[0003]2. Description of the Related Art[0004]Dietary components (supplements), such as vitamins, polyphenols, turmeric, etc. are known to induce various...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/68
CPCA61K31/07A61K31/34A61K31/355A61K31/593A61K36/07C12Q2600/158A61K36/8962C12Q1/6883C12Q1/6886G01N33/5308C12Q2600/136A61K36/746A61K36/00
Inventor MITSUHASHI, MASATO
Owner HITACHI CHEM CO LTD
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